The inducible CRISPR activation(CRISPR-a)system offers unparalleled precision and versatility for regu-lating endogenous genes,making it highly sought after in plant research.In this study,we developed a chem-ically i...The inducible CRISPR activation(CRISPR-a)system offers unparalleled precision and versatility for regu-lating endogenous genes,making it highly sought after in plant research.In this study,we developed a chem-ically inducible CRISPR-a tool for plants called ER-Tag by combining the LexA-VP16-ER inducible system with the SunTag CRISPR-a system.We systematically compared different induction strategies and achieved high efficiency in target gene activation.We demonstrated that guide RNAs can be multiplexed and pooled for large-scale screening of effective morphogenic genes and gene pairs involved in plant regeneration.Further experiments showed that induced activation of these morphogenic genes can accelerate regenera-tion and improve regeneration efficiency in both eudicot and monocot plants,including alfalfa,woodland strawberry,and sheepgrass.Our study expands the CRISPR toolset in plants and provides a powerful new strategy for studying gene function when constitutive expression is not feasible or ideal.展开更多
基金supported by the National Key R&D Program of China (2022YFD1500503 to X.S.)the Key Projects in Science and Technology of Inner Mongolia (2021ZD0031 to X.C.and S.Z.)+6 种基金the National Key Research and Development Program (2022YFF1002802 to X.D.)the Youth Innovation Promotion Association,CAS (Y2022039 to X.D.)the Youth Innovation Promotion Association,CAS (2022096 to S.Z.)the National Science Foundation of China (NSFC32272692 to J.Z.)Project ZR2022QC054 of the Shandong Provincial Natural Science Foundation (to Y.T.)funding from the State Key Laboratory of Protein and Plant Gene Research (to Q.L.).
文摘The inducible CRISPR activation(CRISPR-a)system offers unparalleled precision and versatility for regu-lating endogenous genes,making it highly sought after in plant research.In this study,we developed a chem-ically inducible CRISPR-a tool for plants called ER-Tag by combining the LexA-VP16-ER inducible system with the SunTag CRISPR-a system.We systematically compared different induction strategies and achieved high efficiency in target gene activation.We demonstrated that guide RNAs can be multiplexed and pooled for large-scale screening of effective morphogenic genes and gene pairs involved in plant regeneration.Further experiments showed that induced activation of these morphogenic genes can accelerate regenera-tion and improve regeneration efficiency in both eudicot and monocot plants,including alfalfa,woodland strawberry,and sheepgrass.Our study expands the CRISPR toolset in plants and provides a powerful new strategy for studying gene function when constitutive expression is not feasible or ideal.
